Involvement of Apoptosis in 4-Vinylcyclohexene Diepoxide- Induced Ovotoxicity in Rats
Previous studies have determined that 4-vinylcyclohexene diepoxide (VCD) causes specific destruction of oocytes contained in small pre-antral (primordial and primary) ovarian follicles of Fischer 344 rats following 30 days of daily dosing with VCD. The purposes of this study were to identify the typ...
Gespeichert in:
| Veröffentlicht in: | Toxicology and applied pharmacology 1996-08, Vol.139 (2), p.394-401 |
|---|---|
| Hauptverfasser: | , , , , , |
| Format: | Artikel |
| Sprache: | eng |
| Schlagworte: | |
| Online-Zugang: | Volltext |
| Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
| container_end_page | 401 |
|---|---|
| container_issue | 2 |
| container_start_page | 394 |
| container_title | Toxicology and applied pharmacology |
| container_volume | 139 |
| creator | Springer, L.N. McAsey, M.E. Flaws, J.A. Tilly, J.L. Sipes, I.G. Hoyer, P.B. |
| description | Previous studies have determined that 4-vinylcyclohexene diepoxide (VCD) causes specific destruction of oocytes contained in small pre-antral (primordial and primary) ovarian follicles of Fischer 344 rats following 30 days of daily dosing with VCD. The purposes of this study were to identify the type of VCD-induced cell death occurring in small pre-antral follicles and to determine the earliest time following the onset of dosing when evidence of follicular destruction could first be detected. A significant decrease in the number of oocytes contained in small pre-antral follicles in ovaries of rats after 15 days of daily dosing (ip) with VCD (80 mg/kg) had been observed in preliminary experiments. Therefore, a study was conducted to determine the time of the onset of this follicular destruction by examination of follicular DNA integrity. Female Fischer 344 rats were dosed daily (80 mg/kg, ip) for 6, 8, 10, 12, or 14 days, and ovaries were removed 1, 4, or 24 hr after the final dose. Small pre-antral follicles (25–100 μm) were isolated by gentle dissociation of ovaries with collagenase, and follicles were sorted with micropipets. Genomic DNA was isolated from follicles and radiolabeled with [32P]dideoxy ATP, and the degree of fragmentation quantified by agarose gel electrophoresis and autoradiography. Degradation of DNA was evaluated by32P content in low-molecular-weight fragments ( |
| doi_str_mv | 10.1006/taap.1996.0180 |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_miscellaneous_16058631</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><els_id>S0041008X96901805</els_id><sourcerecordid>16058631</sourcerecordid><originalsourceid>FETCH-LOGICAL-c430t-fa4d25e848dc8a6d1ccc0227b0e079955a9be04f6b1b48b9ba66c3174dce34703</originalsourceid><addsrcrecordid>eNqFkEGLFDEQRoMo6-zq1ZvQB_HWY2WSTifHZVfdgYUFccVbSFeqMdKTtJ3MsPPv7WaGvYmngqr3fRSPsXcc1hxAfSrOjWtujFoD1_CCrTgYVYMQ4iVbAUheA-ifr9llzr8BwEjJL9iF1qB0067Y4zYe0nCgHcVSpb66HtNYUg65CrGS9Y8QjwMecUi_6IkiVbeBxvQUPNXVNvo9kq8eDqnMKwzluIS-uZLfsFe9GzK9Pc8r9vjl8_ebu_r-4ev25vq-Rimg1L2TftOQltqjdspzRITNpu2AoDWmaZzpCGSvOt5J3ZnOKYWCt9IjCdmCuGIfT73jlP7sKRe7CxlpGFyktM-WK2i0Evz_YKNaxeXSuD6BOKWcJ-rtOIWdm46Wg12E20W4XYTbRfgceH9u3nc78s_42fB8_3C-u4xu6CcXMeRnTHDT8HbB9AmjWdch0GQzBoqz3zARFutT-NcHfwGdrZxw</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>15676140</pqid></control><display><type>article</type><title>Involvement of Apoptosis in 4-Vinylcyclohexene Diepoxide- Induced Ovotoxicity in Rats</title><source>MEDLINE</source><source>Elsevier ScienceDirect Journals</source><creator>Springer, L.N. ; McAsey, M.E. ; Flaws, J.A. ; Tilly, J.L. ; Sipes, I.G. ; Hoyer, P.B.</creator><creatorcontrib>Springer, L.N. ; McAsey, M.E. ; Flaws, J.A. ; Tilly, J.L. ; Sipes, I.G. ; Hoyer, P.B.</creatorcontrib><description>Previous studies have determined that 4-vinylcyclohexene diepoxide (VCD) causes specific destruction of oocytes contained in small pre-antral (primordial and primary) ovarian follicles of Fischer 344 rats following 30 days of daily dosing with VCD. The purposes of this study were to identify the type of VCD-induced cell death occurring in small pre-antral follicles and to determine the earliest time following the onset of dosing when evidence of follicular destruction could first be detected. A significant decrease in the number of oocytes contained in small pre-antral follicles in ovaries of rats after 15 days of daily dosing (ip) with VCD (80 mg/kg) had been observed in preliminary experiments. Therefore, a study was conducted to determine the time of the onset of this follicular destruction by examination of follicular DNA integrity. Female Fischer 344 rats were dosed daily (80 mg/kg, ip) for 6, 8, 10, 12, or 14 days, and ovaries were removed 1, 4, or 24 hr after the final dose. Small pre-antral follicles (25–100 μm) were isolated by gentle dissociation of ovaries with collagenase, and follicles were sorted with micropipets. Genomic DNA was isolated from follicles and radiolabeled with [32P]dideoxy ATP, and the degree of fragmentation quantified by agarose gel electrophoresis and autoradiography. Degradation of DNA was evaluated by32P content in low-molecular-weight fragments (<4 kilobase pairs). Degradation of DNA was not observed in follicles collected 24 hr after the final dose on any day. However, a random pattern of DNA degradation was observed, and was significantly greater (p< 0.05) compared with controls, when follicles were collected 4 hr following VCD administration on Days 10 and 12, but not on Days 6 or 8, of dosing. Although not significant, there was also evidence of DNA degradation in dosed animals on Day 14. Histological evaluation of small pre-antral follicles in ovarian sections during the early stages of VCD-induced DNA degradation (Day 10; 4 hr) demonstrated margination of chromatin along the nuclear membrane in oocytes and disruptions in focal contact between granulosa cells and oocytes, both features indicative of apoptosis. Furthermore, there was no sign of ruptured membranes in granulosa cells or oocytes or of an inflammatory response, characteristics of necrosis (pathological cell death). Whereas biochemical and morphological evidence of follicular destruction was seen 4 hr after dosing on Day 10, numbers of oocyte-containing primordial and primary follicles in VCD-treated animals were not different from controls at that time. These results demonstrate that the initial evidence of impending destruction of small pre-antral follicles is first consistently visualized following 10 days of daily dosing with VCD, although a measurable reduction in oocyte numbers has not yet occurred. Despite the fact that internucleosomal cleavage of genomic DNA was not observed, morphological evaluations support that granulosa cells and oocytes in primordial and primary follicles are destroyed via the induction of apoptosis.</description><identifier>ISSN: 0041-008X</identifier><identifier>EISSN: 1096-0333</identifier><identifier>DOI: 10.1006/taap.1996.0180</identifier><identifier>PMID: 8806857</identifier><identifier>CODEN: TXAPA9</identifier><language>eng</language><publisher>San Diego, CA: Elsevier Inc</publisher><subject>Animals ; Apoptosis - drug effects ; Apoptosis - physiology ; Biological and medical sciences ; Carcinogens - administration & dosage ; Carcinogens - toxicity ; Chemical and industrial products toxicology. Toxic occupational diseases ; Cyclohexanes - administration & dosage ; Cyclohexanes - toxicity ; Cyclohexenes ; DNA - analysis ; Female ; Injections, Intraperitoneal ; Medical sciences ; Oocytes - drug effects ; Ovarian Follicle - chemistry ; Ovarian Follicle - drug effects ; Ovary - drug effects ; Ovary - pathology ; Ovary - physiopathology ; Rats ; Rats, Inbred F344 ; Toxicology ; Various organic compounds ; Vinyl Compounds - administration & dosage ; Vinyl Compounds - toxicity</subject><ispartof>Toxicology and applied pharmacology, 1996-08, Vol.139 (2), p.394-401</ispartof><rights>1996 Academic Press</rights><rights>1996 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c430t-fa4d25e848dc8a6d1ccc0227b0e079955a9be04f6b1b48b9ba66c3174dce34703</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1006/taap.1996.0180$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,777,781,3537,27905,27906,45976</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=3195177$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/8806857$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Springer, L.N.</creatorcontrib><creatorcontrib>McAsey, M.E.</creatorcontrib><creatorcontrib>Flaws, J.A.</creatorcontrib><creatorcontrib>Tilly, J.L.</creatorcontrib><creatorcontrib>Sipes, I.G.</creatorcontrib><creatorcontrib>Hoyer, P.B.</creatorcontrib><title>Involvement of Apoptosis in 4-Vinylcyclohexene Diepoxide- Induced Ovotoxicity in Rats</title><title>Toxicology and applied pharmacology</title><addtitle>Toxicol Appl Pharmacol</addtitle><description>Previous studies have determined that 4-vinylcyclohexene diepoxide (VCD) causes specific destruction of oocytes contained in small pre-antral (primordial and primary) ovarian follicles of Fischer 344 rats following 30 days of daily dosing with VCD. The purposes of this study were to identify the type of VCD-induced cell death occurring in small pre-antral follicles and to determine the earliest time following the onset of dosing when evidence of follicular destruction could first be detected. A significant decrease in the number of oocytes contained in small pre-antral follicles in ovaries of rats after 15 days of daily dosing (ip) with VCD (80 mg/kg) had been observed in preliminary experiments. Therefore, a study was conducted to determine the time of the onset of this follicular destruction by examination of follicular DNA integrity. Female Fischer 344 rats were dosed daily (80 mg/kg, ip) for 6, 8, 10, 12, or 14 days, and ovaries were removed 1, 4, or 24 hr after the final dose. Small pre-antral follicles (25–100 μm) were isolated by gentle dissociation of ovaries with collagenase, and follicles were sorted with micropipets. Genomic DNA was isolated from follicles and radiolabeled with [32P]dideoxy ATP, and the degree of fragmentation quantified by agarose gel electrophoresis and autoradiography. Degradation of DNA was evaluated by32P content in low-molecular-weight fragments (<4 kilobase pairs). Degradation of DNA was not observed in follicles collected 24 hr after the final dose on any day. However, a random pattern of DNA degradation was observed, and was significantly greater (p< 0.05) compared with controls, when follicles were collected 4 hr following VCD administration on Days 10 and 12, but not on Days 6 or 8, of dosing. Although not significant, there was also evidence of DNA degradation in dosed animals on Day 14. Histological evaluation of small pre-antral follicles in ovarian sections during the early stages of VCD-induced DNA degradation (Day 10; 4 hr) demonstrated margination of chromatin along the nuclear membrane in oocytes and disruptions in focal contact between granulosa cells and oocytes, both features indicative of apoptosis. Furthermore, there was no sign of ruptured membranes in granulosa cells or oocytes or of an inflammatory response, characteristics of necrosis (pathological cell death). Whereas biochemical and morphological evidence of follicular destruction was seen 4 hr after dosing on Day 10, numbers of oocyte-containing primordial and primary follicles in VCD-treated animals were not different from controls at that time. These results demonstrate that the initial evidence of impending destruction of small pre-antral follicles is first consistently visualized following 10 days of daily dosing with VCD, although a measurable reduction in oocyte numbers has not yet occurred. Despite the fact that internucleosomal cleavage of genomic DNA was not observed, morphological evaluations support that granulosa cells and oocytes in primordial and primary follicles are destroyed via the induction of apoptosis.</description><subject>Animals</subject><subject>Apoptosis - drug effects</subject><subject>Apoptosis - physiology</subject><subject>Biological and medical sciences</subject><subject>Carcinogens - administration & dosage</subject><subject>Carcinogens - toxicity</subject><subject>Chemical and industrial products toxicology. Toxic occupational diseases</subject><subject>Cyclohexanes - administration & dosage</subject><subject>Cyclohexanes - toxicity</subject><subject>Cyclohexenes</subject><subject>DNA - analysis</subject><subject>Female</subject><subject>Injections, Intraperitoneal</subject><subject>Medical sciences</subject><subject>Oocytes - drug effects</subject><subject>Ovarian Follicle - chemistry</subject><subject>Ovarian Follicle - drug effects</subject><subject>Ovary - drug effects</subject><subject>Ovary - pathology</subject><subject>Ovary - physiopathology</subject><subject>Rats</subject><subject>Rats, Inbred F344</subject><subject>Toxicology</subject><subject>Various organic compounds</subject><subject>Vinyl Compounds - administration & dosage</subject><subject>Vinyl Compounds - toxicity</subject><issn>0041-008X</issn><issn>1096-0333</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1996</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkEGLFDEQRoMo6-zq1ZvQB_HWY2WSTifHZVfdgYUFccVbSFeqMdKTtJ3MsPPv7WaGvYmngqr3fRSPsXcc1hxAfSrOjWtujFoD1_CCrTgYVYMQ4iVbAUheA-ifr9llzr8BwEjJL9iF1qB0067Y4zYe0nCgHcVSpb66HtNYUg65CrGS9Y8QjwMecUi_6IkiVbeBxvQUPNXVNvo9kq8eDqnMKwzluIS-uZLfsFe9GzK9Pc8r9vjl8_ebu_r-4ev25vq-Rimg1L2TftOQltqjdspzRITNpu2AoDWmaZzpCGSvOt5J3ZnOKYWCt9IjCdmCuGIfT73jlP7sKRe7CxlpGFyktM-WK2i0Evz_YKNaxeXSuD6BOKWcJ-rtOIWdm46Wg12E20W4XYTbRfgceH9u3nc78s_42fB8_3C-u4xu6CcXMeRnTHDT8HbB9AmjWdch0GQzBoqz3zARFutT-NcHfwGdrZxw</recordid><startdate>19960801</startdate><enddate>19960801</enddate><creator>Springer, L.N.</creator><creator>McAsey, M.E.</creator><creator>Flaws, J.A.</creator><creator>Tilly, J.L.</creator><creator>Sipes, I.G.</creator><creator>Hoyer, P.B.</creator><general>Elsevier Inc</general><general>Elsevier</general><scope>IQODW</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7U7</scope><scope>C1K</scope></search><sort><creationdate>19960801</creationdate><title>Involvement of Apoptosis in 4-Vinylcyclohexene Diepoxide- Induced Ovotoxicity in Rats</title><author>Springer, L.N. ; McAsey, M.E. ; Flaws, J.A. ; Tilly, J.L. ; Sipes, I.G. ; Hoyer, P.B.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c430t-fa4d25e848dc8a6d1ccc0227b0e079955a9be04f6b1b48b9ba66c3174dce34703</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1996</creationdate><topic>Animals</topic><topic>Apoptosis - drug effects</topic><topic>Apoptosis - physiology</topic><topic>Biological and medical sciences</topic><topic>Carcinogens - administration & dosage</topic><topic>Carcinogens - toxicity</topic><topic>Chemical and industrial products toxicology. Toxic occupational diseases</topic><topic>Cyclohexanes - administration & dosage</topic><topic>Cyclohexanes - toxicity</topic><topic>Cyclohexenes</topic><topic>DNA - analysis</topic><topic>Female</topic><topic>Injections, Intraperitoneal</topic><topic>Medical sciences</topic><topic>Oocytes - drug effects</topic><topic>Ovarian Follicle - chemistry</topic><topic>Ovarian Follicle - drug effects</topic><topic>Ovary - drug effects</topic><topic>Ovary - pathology</topic><topic>Ovary - physiopathology</topic><topic>Rats</topic><topic>Rats, Inbred F344</topic><topic>Toxicology</topic><topic>Various organic compounds</topic><topic>Vinyl Compounds - administration & dosage</topic><topic>Vinyl Compounds - toxicity</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Springer, L.N.</creatorcontrib><creatorcontrib>McAsey, M.E.</creatorcontrib><creatorcontrib>Flaws, J.A.</creatorcontrib><creatorcontrib>Tilly, J.L.</creatorcontrib><creatorcontrib>Sipes, I.G.</creatorcontrib><creatorcontrib>Hoyer, P.B.</creatorcontrib><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Toxicology Abstracts</collection><collection>Environmental Sciences and Pollution Management</collection><jtitle>Toxicology and applied pharmacology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Springer, L.N.</au><au>McAsey, M.E.</au><au>Flaws, J.A.</au><au>Tilly, J.L.</au><au>Sipes, I.G.</au><au>Hoyer, P.B.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Involvement of Apoptosis in 4-Vinylcyclohexene Diepoxide- Induced Ovotoxicity in Rats</atitle><jtitle>Toxicology and applied pharmacology</jtitle><addtitle>Toxicol Appl Pharmacol</addtitle><date>1996-08-01</date><risdate>1996</risdate><volume>139</volume><issue>2</issue><spage>394</spage><epage>401</epage><pages>394-401</pages><issn>0041-008X</issn><eissn>1096-0333</eissn><coden>TXAPA9</coden><abstract>Previous studies have determined that 4-vinylcyclohexene diepoxide (VCD) causes specific destruction of oocytes contained in small pre-antral (primordial and primary) ovarian follicles of Fischer 344 rats following 30 days of daily dosing with VCD. The purposes of this study were to identify the type of VCD-induced cell death occurring in small pre-antral follicles and to determine the earliest time following the onset of dosing when evidence of follicular destruction could first be detected. A significant decrease in the number of oocytes contained in small pre-antral follicles in ovaries of rats after 15 days of daily dosing (ip) with VCD (80 mg/kg) had been observed in preliminary experiments. Therefore, a study was conducted to determine the time of the onset of this follicular destruction by examination of follicular DNA integrity. Female Fischer 344 rats were dosed daily (80 mg/kg, ip) for 6, 8, 10, 12, or 14 days, and ovaries were removed 1, 4, or 24 hr after the final dose. Small pre-antral follicles (25–100 μm) were isolated by gentle dissociation of ovaries with collagenase, and follicles were sorted with micropipets. Genomic DNA was isolated from follicles and radiolabeled with [32P]dideoxy ATP, and the degree of fragmentation quantified by agarose gel electrophoresis and autoradiography. Degradation of DNA was evaluated by32P content in low-molecular-weight fragments (<4 kilobase pairs). Degradation of DNA was not observed in follicles collected 24 hr after the final dose on any day. However, a random pattern of DNA degradation was observed, and was significantly greater (p< 0.05) compared with controls, when follicles were collected 4 hr following VCD administration on Days 10 and 12, but not on Days 6 or 8, of dosing. Although not significant, there was also evidence of DNA degradation in dosed animals on Day 14. Histological evaluation of small pre-antral follicles in ovarian sections during the early stages of VCD-induced DNA degradation (Day 10; 4 hr) demonstrated margination of chromatin along the nuclear membrane in oocytes and disruptions in focal contact between granulosa cells and oocytes, both features indicative of apoptosis. Furthermore, there was no sign of ruptured membranes in granulosa cells or oocytes or of an inflammatory response, characteristics of necrosis (pathological cell death). Whereas biochemical and morphological evidence of follicular destruction was seen 4 hr after dosing on Day 10, numbers of oocyte-containing primordial and primary follicles in VCD-treated animals were not different from controls at that time. These results demonstrate that the initial evidence of impending destruction of small pre-antral follicles is first consistently visualized following 10 days of daily dosing with VCD, although a measurable reduction in oocyte numbers has not yet occurred. Despite the fact that internucleosomal cleavage of genomic DNA was not observed, morphological evaluations support that granulosa cells and oocytes in primordial and primary follicles are destroyed via the induction of apoptosis.</abstract><cop>San Diego, CA</cop><pub>Elsevier Inc</pub><pmid>8806857</pmid><doi>10.1006/taap.1996.0180</doi><tpages>8</tpages></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0041-008X |
| ispartof | Toxicology and applied pharmacology, 1996-08, Vol.139 (2), p.394-401 |
| issn | 0041-008X 1096-0333 |
| language | eng |
| recordid | cdi_proquest_miscellaneous_16058631 |
| source | MEDLINE; Elsevier ScienceDirect Journals |
| subjects | Animals Apoptosis - drug effects Apoptosis - physiology Biological and medical sciences Carcinogens - administration & dosage Carcinogens - toxicity Chemical and industrial products toxicology. Toxic occupational diseases Cyclohexanes - administration & dosage Cyclohexanes - toxicity Cyclohexenes DNA - analysis Female Injections, Intraperitoneal Medical sciences Oocytes - drug effects Ovarian Follicle - chemistry Ovarian Follicle - drug effects Ovary - drug effects Ovary - pathology Ovary - physiopathology Rats Rats, Inbred F344 Toxicology Various organic compounds Vinyl Compounds - administration & dosage Vinyl Compounds - toxicity |
| title | Involvement of Apoptosis in 4-Vinylcyclohexene Diepoxide- Induced Ovotoxicity in Rats |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-01-20T10%3A13%3A11IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_cross&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Involvement%20of%20Apoptosis%20in%204-Vinylcyclohexene%20Diepoxide-%20Induced%20Ovotoxicity%20in%20Rats&rft.jtitle=Toxicology%20and%20applied%20pharmacology&rft.au=Springer,%20L.N.&rft.date=1996-08-01&rft.volume=139&rft.issue=2&rft.spage=394&rft.epage=401&rft.pages=394-401&rft.issn=0041-008X&rft.eissn=1096-0333&rft.coden=TXAPA9&rft_id=info:doi/10.1006/taap.1996.0180&rft_dat=%3Cproquest_cross%3E16058631%3C/proquest_cross%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=15676140&rft_id=info:pmid/8806857&rft_els_id=S0041008X96901805&rfr_iscdi=true |